Multipotent adult stem cells, sources thereof, methods of obtaining and maintaining same, methods of differentiation thereof, methods of use thereof and cells derived thereof

Abstract

Methods and compositions are provided for circularizing target sequences in a sample. In particular, ligation oligonucleotides are employed to selectively hybridize with the target such that the target can be ligated into a closed circular target. Rolling circle amplification can then be performed directly on the target sequence for subsequent detection and analysis.

Description

RELATED CASES [0001] This application claims the benefit of U.S. Provisional Application No. 60 / 343,386, filed Oct. 25, 2001, U.S. Provisional Application No. 60 / 310,625, filed Aug. 7, 2001, U.S. Provisional Application No. 60 / 269,062, filed Feb. 15, 2001, U.S. Provisional Application No. 60 / 268,786, filed Feb. 14, 2001, which are hereby incorporated by reference for all purposes. Applicants also claim priority of WO 01 / 11011, 60 / 147,324 and 60 / 164,650 and these applications are hereby incorporated by reference into this text; any teachings therein may be used in the practice of this invention. The present application is a continuation-in-part of 15′ WO 01 / 11011, which is attached herein at Appendix 1 and is part of the present application. Documents incorporated by reference into this text are not admitted to be prior art.FIELD OF THE INVENTION [0002] The present invention relates generally to mammalian multipotent adult stem cells (MASC), and more specifically to methods for obtai...

AI Technical Summary

Benefits of technology

[0040] An important application of this technology is the method of treating a patient by administering a therapeutically effective amount of MASC or their progeny. The progeny can either have the capacity to be further differentiated, or can be terminally differentiated. An unexpected benefit of this approach is that the need for pretreatment and / or post treatment of the patient with irradiation, chemotherapy, immunosuppressive agents or other drugs or treatments is reduced or eliminated. The induction of tolerance before or during treatment is also not required.

[0047] In another preferred embodiment, the MASC or their progeny enhance angiogenesis.

Problems solved by technology

However, there are at least two major problems associated with organ and tissue transplantation.

First, there is a shortage of donor organs and tissues.

The second major problem is the potential incompatibility of the transplanted tissue with the immune system of the recipient.

Because the donated organ or tissue is recognized by the host immune system as foreign, immunosuppressive medications must be provided to the patient at a significant cost-both financially and physically.

Unfortunately, xenotransplantation does not overcome the problem of tissue incompatibility, but instead exacerbates it.

Furthermore, according to the Centers for Disease Control, there is evidence that damaging viruses cross species barriers.

Stem cells which differentiate only to form cells of hematopoietic lineage, however, are unable to provide a source of cells for repair of other damaged tissues, for example, heart or lung tissue damaged by high-dose chemotherapeutic agents.

The most commonly used source of NSC is allogeneic fetal brain, which poses both immunological and ethical problems.

As it is not known whether pre-existing neural pathology will affect the ability of NSC to be cultured and induced to differentiate into neuronal and glial cells ex vivo, and because additional surgery in an already diseased brain may aggravate the underlying disease, this approach is less attractive.

This cell is capable of differentiating to form a number of cell types of mesenchyrnal origin, but is apparently limited in differentiation potential to cells of the mesenchymal lineage, as the team who isolated it noted that hematopoietic cells were never identified in the expanded cultures (Pittenger, el al., 1999).

In addition, the degree of telomerase activity in tissue specific or lineage comitted stem cells has not been fully explored, in part because large numbers of highly enriched populations of these cells are difficult to obtain.

However, the true nature of these types of cells has not been fully discerned.

Also, stem cells isolated from a given organ may not necessarily be a lineage committed cell.

Indeed most investigators did not identify the phenotype of the initiating cell.

Thus, although ES cell research provides a promising alternative solution to the problem of a limited supply of organs for transplantation, the problems and risks associated with the need for immunosuppression to sustain transplantation of heterologous cells or tissue would remain.

The greatest disadvantage associated with the use of autologous stem cells for tissue transplant thus far lies in their relatively limited differentiation potential.

A number of stem cells have been isolated from fully-developed organisms, particularly humans, but these cells, although reported to be multipotent, have demonstrated limited potential to differentiate to multiple cell types.

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